Tomato hybrid Onuba

ABSTRACT

The present invention provides tomato hybrid Onuba and plant parts, seed, and tissue culture therefrom. The invention also provides methods for producing a tomato plant by crossing the tomato plants of the invention with themselves or another tomato plant. The invention also provides tomato plants produced from such a crossing as well as plant parts, seed, and tissue culture therefrom.

FIELD OF THE INVENTION

This invention is in the field of tomato plants.

BACKGROUND OF THE INVENTION

Tomato is an important global crop. Several plant species associatedwith the family Solanaceae have been familiar to mankind since ancienttimes and are of great agricultural importance. The genus Solanum is amember of the Solanaceae family and includes the domesticated tomato S.lycopersicum (also known as Lycopersicum esculentum). Tomato isgenerally adapted to warm summer growing conditions but can also begrown in heated greenhouses under winter conditions. The introduction ofhybrid tomato cultivars in the 1950s provided a magnitude of benefitsincluding increased yield, better holding ability, adaptation toexpanded growing seasons through the use of protected cultivation andimproved disease resistances, which resulted in large-scale productionof tomato as a commercial crop.

Tomato breeders make continual improvements in hybrid tomato yield,horticultural characteristics, and consumer quality traits. Thus, thereis an ongoing need for improved tomato hybrid varieties and parentallines.

SUMMARY OF THE INVENTION

According to the invention, there is provided a novel hybrid tomatovariety, designated Onuba, also known as 603345.

The invention also encompasses the seeds of tomato hybrid 603345, theplants of tomato hybrid 603345, plant parts of the tomato hybrid 603345(including fruit, seed, gametes, rootstock, scions, shoots), methods ofproducing seed from tomato hybrid 603345, and methods for producing atomato plant by crossing the tomato hybrid 603345 with itself or anothertomato plant, methods for producing a tomato plant containing in itsgenetic material one or more transgenes, and the transgenic tomatoplants produced by that method. The invention also relates to methodsfor producing other tomato plants derived from tomato hybrid 603345 andto tomato plants, parts thereof and seed produced by the use of thosemethods. The present invention further relates to hybrid tomato seedsand plants (and parts thereof including fruit) produced by crossingtomato hybrid 603345 with another tomato plant.

The invention further contemplates grafted tomato plants and methods ofproducing a grafted tomato plant, where tomato hybrid 603345 can be usedas either the rootstock or the scion.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of tomato hybrid 603345. In embodiments, thetissue culture is capable of regenerating plants having all oressentially all of the physiological and morphological characteristicsof the foregoing tomato plant and/or of regenerating plants having thesame or substantially the same genotype as the foregoing tomato plant.In embodiments, the regenerated plant is a diploid plant. In exemplaryembodiments, the regenerable cells in such tissue cultures aremeristematic cells, cotyledons, hypocotyl, leaves, pollen, embryos,roots, root tips, anthers, pistils, ovules, shoots, stems, petiole,pith, flowers, capsules and/or seeds as well as callus and/orprotoplasts derived from any of the foregoing. Still further, thepresent invention provides tomato plants regenerated from the tissuecultures of the invention.

As a further aspect, the invention provides a method of producing tomatoseed, the method comprising crossing a plant of tomato hybrid 603345with itself or a second tomato plant, cultivating the plant to maturity,and allowing seed to form. Optionally, the method further comprisescollecting the seed.

Another aspect of the invention provides methods for producing hybridsand other tomato plants derived from tomato hybrid 603345. Tomato plantsderived by the use of these methods are also part of the invention aswell as plant parts, seed, gametes and tissue culture from such hybridor derived tomato plants.

In representative embodiments, a tomato plant derived from tomato hybrid603345 comprises cells comprising at least one set of chromosomesderived from tomato hybrid 603345.

In embodiments, a tomato plant or population of tomato plants derivedfrom tomato hybrid 603345 comprises, on average, at least 6.25%, 12.5%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%,96%, 97%, 98% or 99% of its alleles (i.e., theoretical allelic content;TAC) from tomato hybrid 603345, e.g., at least about 6.25%, 12.5%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%,97%, 98% or 99% of the genetic complement of tomato hybrid 603345,respectively, and optionally may be the result of a breeding processcomprising one or two breeding crosses and one or more of selfing,sibbing, backcrossing and/or double haploid techniques in anycombination and any order. In embodiments, the breeding process does notinclude a breeding cross, and comprises selfing, sibbing, backcrossingand or double haploid technology. In embodiments, the tomato plantderived from tomato hybrid 603345 is one, two, three, four, five or morebreeding crosses removed from tomato hybrid 603345, respectively.

In embodiments, a hybrid or derived plant from tomato hybrid 603345comprises a desired added trait(s). In representative embodiments, atomato plant derived from tomato hybrid 603345 comprises all of themorphological and physiological characteristics of tomato hybrid 603345(e.g., as described in Table 1), respectively, with the addition of thedesired added trait(s). In embodiments, the tomato plant derived fromtomato hybrid 603345 comprises essentially all of the morphological andphysiological characteristics of tomato hybrid 603345 (e.g., asdescribed in Table 1), respectively, with the addition of a desiredadded trait(s).

The invention also relates to methods for producing a tomato plantcomprising in its genetic material one or more transgenes and to thetransgenic tomato plant produced by those methods (and progeny tomatoplants comprising the transgene). Also provided are plant parts, seedand tissue culture from such transgenic tomato plants, optionallywherein one or more cells in the plant part, seed, or tissue culturecomprises the transgene. The transgene can be introduced via planttransformation and/or breeding techniques (e.g., once planttransformation of a precursor plant has been achieved, it can then betransferred to other plants via conventional breeding).

In another aspect, the present invention provides for single locusconverted plants of tomato hybrid 603345. Plant parts, seed, and tissueculture from such single locus converted plants are also contemplated bythe present invention. The single locus may be a dominant or recessiveallele. The single locus may be a naturally occurring tomato locus(e.g., introduced via traditional breeding or gene editing techniques),a transgene introduced into tomato through genetic engineeringtechniques (of the plant or a precursor thereof), or a non-naturallyoccurring locus (e.g., produced by mutagenesis or gene editing).

The invention further provides methods for developing tomato plants in atomato plant breeding program using plant breeding techniques including,for example, recurrent selection, backcrossing, pedigree breeding,double haploid techniques, restriction fragment length polymorphismenhanced selection, genetic marker enhanced selection, gene editingand/or genetic transformation. Seeds, tomato plants, and parts thereof,produced by such breeding methods are also part of the invention.

The invention also provides methods of multiplication or propagation oftomato plants of the invention, which can be accomplished using anymethod known in the art, for example, via vegetative propagation and/orseed.

Additional aspects of the invention include harvested products andprocessed products from the tomato plants of the invention. A harvestedproduct can be a whole plant or any plant part, as described herein.Thus, in some embodiments, a non-limiting example of a harvested productincludes a seed, a fruit (e.g., including the flesh), a rootstock, ascion and/or a shoot.

In representative embodiments, a processed product includes, but is notlimited to: cut, sliced, ground, dried, pureed, canned, jarred, washed,packaged, frozen, seeded, peeled and/or heated fruit or fruit flesh ofthe tomato plants of the invention, or any other part thereof, includingpuree, paste, sauce and salsa made from fruits or fruit flesh.

The seed of the invention can optionally be provided as an essentiallyhomogenous population of seed of a single plant or cultivar. Essentiallyhomogenous populations of seed are generally free from substantialnumbers of other seed, e.g., at least about 90%, 95%, 96%, 97%, 98% or99% pure.

In representative embodiments, the invention provides a seed of tomatohybrid 603345.

As a further aspect, the invention provides a plant of tomato hybrid603345.

As an additional aspect, the invention provides a tomato plant, or apart thereof, having all or essentially all of the physiological andmorphological characteristics of a plant of tomato hybrid 603345.

As another aspect, the invention provides fruit and fruit flesh of thetomato plants of the invention and processed products from the fruit orfruit flesh of the inventive tomato plants.

As still another aspect, the invention provides a method of producingtomato seed, the method comprising crossing a tomato plant of theinvention with itself or a second tomato plant to produce progeny seed.In embodiments, the method is practiced to self the plant of theinvention. The invention also provides seed produced by this method andplants, and parts thereof including fruit, produced by growing the seed.

As yet a further aspect, the invention provides a method for producing aseed of a tomato plant derived from tomato hybrid 603345, the methodcomprising: (a) crossing a tomato plant of tomato hybrid 603345 with asecond tomato plant; and (b) allowing seed of a tomato plant derivedfrom tomato hybrid 603345 to form. In embodiments, the method furthercomprises: (c) growing a plant from the seed derived from tomato hybrid603345 of step (b); (d) selfing the plant of step (c) or crossing it toa second tomato plant to form additional tomato seed derived from tomatohybrid 603345, and (e) optionally repeating steps (c) and (d) one ormore times to generate further derived tomato seed from tomato hybrid603345, wherein in step (c) a plant is grown from the additional tomatoseed of step (d) in place of growing a plant from the seed of step (b).In embodiments, the method comprises: (e) repeating steps (c) and (d)one or more times (e.g., three or more, four or more, five or more, sixor more, one to three, one to five, one to six, one to seven, one toten, three to five, three to six, three to seven, three to eight orthree to ten times) to generate further derived tomato seed. As anotheroption, the method can comprise cultivating the plant to maturity andcollecting the seed. The invention also provides seed produced by thesemethods and tomato plants produced by growing the seed.

Still further, as another aspect, the invention provides a method ofvegetatively propagating a plant of tomato hybrid 603345, e.g., viashoot proliferation and then rooting in tissue culture. In anon-limiting example, the method comprises: (a) collecting tissuecapable of being propagated from a plant of tomato hybrid 603345; (b)cultivating the tissue to obtain proliferated shoots; and (c) rootingthe proliferated shoots to obtain rooted plantlets. Optionally, theinvention further comprises growing plants from the rooted plantlets.The invention also encompasses the plantlets and plants produced bythese methods, as well as seed and fruits produced therefrom.

As a further aspect, the invention provides a method of producing agrafted tomato plant, wherein hybrid 603345 is used as either therootstock or the scion.

As an additional aspect, the invention provides a method of introducinga desired added trait into tomato hybrid 603345, the method comprisingconverting a parent line (male and/or female) of tomato hybrid 603345 tocomprise the desired added trait. In embodiments in which the desiredadded trait is recessive, both parent lines are converted.

In representative embodiments, the invention also provides a method ofproducing a plant of tomato hybrid 603345 comprising a desired addedtrait, the method comprising introducing a transgene conferring thedesired trait into a plant of tomato hybrid 603345. In embodiments, thetransgene can be introduced by transformation methods (e.g., geneticengineering) or traditional breeding techniques into one or both parentlines or a precursor thereof.

The invention also provides tomato plants produced by the methods of theinvention, wherein the tomato plant has the desired added trait as wellas seed and fruits from such tomato plants. The invention also providesseed that produces the plants derived from hybrid 603345 and comprisinga desired added trait.

According to the foregoing methods, the desired added trait can be anysuitable trait known in the art including without limitation: malesterility, male fertility, herbicide resistance, pest (e.g., insectand/or nematode) resistance, modified fatty acid metabolism, modifiedcarbohydrate metabolism, disease resistance (e.g., against bacterial,fungal and/or viral disease), abiotic stress tolerance, enhancednutritional quality (e.g., increased flavonoid content of the fruit),improved appearance (e.g., external fruit color and/or fruit fleshcolor), increased fruit sweetness, increased fruit flavor, improvedfruit ripening, improved texture or taste, improved fruit yield,improved seed yield, improved seedling vigor, improved seed germination,industrial usage, or any combination thereof.

In representative embodiments, a transgene conferring herbicideresistance confers resistance to glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine,benzonitrile, or any combination thereof.

In representative embodiments, a transgene conferring pest resistance(e.g., insect and/or nematode resistance) encodes a Bacillusthuringiensis endotoxin.

In representative embodiments, transgenic plants, transformed plants(e.g., using genetic engineering techniques), gene edited plants, singlelocus converted plants, hybrid plants and tomato plants derived fromtomato hybrid 603345 are characterized by 1, 2, 3, 4 or more of thecharacteristics of tomato hybrid 603345 as described herein, e.g.,produces mini-clusters of small mature fruit with a pale red color, around shape, and/or a high fructose content. In embodiments, transgenicplants, transformed plants, gene edited plants, hybrid plants and tomatoplants derived from tomato hybrid 603345 comprise essentially all of themorphological and physiological characteristics of tomato hybrid 603345(for example, as described in Table 1), respectively, or even all of themorphological and physiological characteristics of tomato hybrid 603345,so that said plants are not significantly different for said traits thantomato hybrid 603345, respectively, as determined at the 5% significancelevel when grown in the same environmental conditions; optionally, withthe presence of one or more desired additional traits (e.g., asdescribed above).

In one embodiment, the present invention discloses a method of producingseeds of tomato hybrid 603345 comprising growing a plant of tomatohybrid 603345; allowing pollination of said plants, for example,open-pollination of said plants in an isolated plot or field; andharvesting seeds from said plants. In one embodiment, the method furthercomprises washing and drying said seed.

The invention further provides a method of developing a tomato variety(e.g., a diploid tomato variety) in a tomato plant breeding programusing plant breeding techniques, which include employing a tomato plant,or a part thereof, as a source of plant breeding material, the methodcomprising: (a) obtaining the tomato plant, or a part thereof, of hybrid603345 as a source of breeding material; and (b) applying plant breedingtechniques.

The invention also encompasses plant parts, plant material, pollen,ovules, leaves, fruit and seed from the tomato plants of the invention.The invention also provides seed that produces the tomato plants of theinvention. Also provided is a tissue culture of regenerable cells fromthe tomato plants of the invention, where optionally, the regenerablecells are: (a) embryos, meristem, leaves, pollen, cotyledons,hypocotyls, roots, root tips, anthers, flowers, pistils, ovules, seed,shoots, stems, stalks, petioles, pith and/or capsules; or (b) callus orprotoplasts derived from the cells of (a). Further provided are tomatoplants regenerated from a tissue culture of the invention.

In still yet another aspect, the invention provides a method ofdetermining a genetic characteristic of tomato hybrid 603345 or aprogeny thereof, e.g., a method of determining a genotype of tomatohybrid 603345 or a progeny thereof using molecular genetic techniques.In embodiments, the method comprises detecting in the genome of a tomatohybrid 603345 plant, or a progeny plant thereof, at least a firstpolymorphism, e.g., comprises nucleic acid amplification and/or nucleicacid sequencing. To illustrate, in embodiments, the method comprisesobtaining a sample of nucleic acids from the plant and detecting atleast a first polymorphism in the nucleic acid sample (e.g., using oneor more molecular markers). Optionally, the method may comprisedetecting a plurality of polymorphisms (e.g., two or more, three ormore, four or more, five or more, six or more, eight or more or ten ormore polymorphisms, etc.) in the genome of the plant. In representativeembodiments, the method further comprises storing the results of thestep of detecting the polymorphism(s) on a computer readable medium. Theinvention further provides a computer readable medium produced by such amethod.

In addition to the exemplary aspects and embodiments described above,the invention is described in more detail in the description of theinvention set forth below.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based, in part, on the development of a noveltomato hybrid 603345. Among other characteristics, when grown understandard horticultural conditions (e.g., greenhouse), tomato hybrid603345 is characterized by mini clusters of small mature fruit with apale red color, a round shape, average weight of 24 grams, and/or a highfructose content.

It should be appreciated that the invention can be embodied in differentforms and should not be construed as limited to the embodiments setforth herein. Rather, these embodiments are provided so that thisdisclosure will be thorough and complete, and will fully convey thescope of the invention to those skilled in the art.

Unless the context indicates otherwise, it is specifically intended thatthe various features and embodiments of the invention described hereincan be used in any combination.

Moreover, the present invention also contemplates that in someembodiments of the invention, any feature or combination of features setforth herein can be excluded or omitted. To illustrate, if thespecification states that a composition comprises components A, B and C,it is specifically intended that any of A, B or C, or a combinationthereof, can be omitted and disclaimed singularly or in any combination.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. The terminology used in thedescription of the invention herein is for the purpose of describingparticular embodiments only and is not intended to be limiting of theinvention.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

As used in the description of the invention and the appended claims, thesingular forms “a,” “an” and “the” are intended to include the pluralforms as well, unless the context clearly indicates otherwise.

As used herein, “and/or” refers to and encompasses any and all possiblecombinations of one or more of the associated listed items, as well asthe lack of combinations when interpreted in the alternative (“or”).

The term “about,” as used herein when referring to a measurable valuesuch as a dosage or time period and the like, is meant to encompassvariations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of thespecified amount.

The term “comprise,” “comprises” and “comprising” as used herein,specify the presence of the stated features, integers, steps,operations, elements, and/or components, but do not preclude thepresence or addition of one or more other features, integers, steps,operations, elements, components, and/or groups thereof.

As used herein, the transitional phrase “consisting essentially of”means that the scope of a claim is to be interpreted to encompass thespecified materials or steps recited in the claim “and those that do notmaterially affect the basic and novel characteristic(s)” of the claimedinvention. See, In re Herz, 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463(CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus,the term “consisting essentially of” when used in a claim or thedescription of this invention is not intended to be interpreted to beequivalent to “comprising.”

“Added trait”: As used herein, a desired “added trait” or “additionaltrait” can be any trait that confers a desired characteristic to theplant, and may be introduced by any method known in the art, e.g.,conventional breeding (for example, backcrossing), gene editing,mutagenesis, or genetic transformation techniques. Examples of addedtraits include without limitation: male sterility, male fertility,herbicide resistance, pest (e.g., insect and/or nematode) resistance,modified fatty acid metabolism, modified carbohydrate metabolism,disease resistance (e.g., against bacterial, fungal and/or viraldisease), abiotic stress tolerance, enhanced nutritional quality (e.g.,increased flavonoid content of the fruit), improved appearance (e.g.,external fruit color and/or fruit flesh color), increased fruitsweetness, increased fruit flavor, improved fruit ripening, improvedtexture or taste, improved fruit yield, improved seed yield, improvedseedling vigor, improved seed germination, industrial usage, geneediting machinery, or any combination thereof. The trait can be encodedby a DNA sequence (e.g., native tomato sequence, induced or naturallyoccurring mutation, edited native sequence, or a transgene) or canresult from expression of a functional non-translated RNA (e.g., RNAinterference).

“Allele”. An allele is any of one or more alternative forms of a gene,all of which relate to a trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy the corresponding locuson a pair of homologous chromosomes.

“Backcrossing”. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents (the “recurrent”parent), for example, a first generation hybrid F₁ with one of theparental genotype of the F₁ hybrid.

“Cotyledon”. One of the first leaves of the embryo of a seed plant;typically one or more in monocotyledons, two in dicotyledons, and two ormore in gymnosperms.

“Determinate”: Determinate tomato plants tend to be compact and bushy inform, and the plant produces all of its fruit simultaneously within ashort period of time (e.g., one to three weeks), and then fruitproduction stops.

“Double haploid line”. A stable inbred line achieved by doubling thechromosomes of a haploid line, e.g., from anther culture. For example,some pollen grains (haploid) cultivated under specific conditionsdevelop plantlets containing 1 n chromosomes. The chromosomes in theseplantlets are then induced to “double” (e.g., using chemical means)resulting in cells containing 2n chromosomes. The progeny of theseplantlets are termed “double haploid” and are essentially notsegregating any more (e.g., are stable). The term “double haploid” isused interchangeably herein with “dihaploid.”

“Essentially all of the physiological and morphologicalcharacteristics”. A plant having “essentially all the physiological andmorphological characteristics” (and similar phrases) means a planthaving all of the desired physiological and morphologicalcharacteristics of hybrid 603345, except for the characteristic(s)derived from a converted locus/loci (e.g., a single converted locus),for example, introduced via backcrossing to hybrid 603345, a modifiedgene(s) resulting from genome editing techniques, an introducedtransgene (i.e., introduced via genetic transformation techniques) ormutation, when both plants are grown under the same environmentalconditions. In embodiments, a plant having “essentially all of thephysiological and morphological characteristics” means a plant havingall of the characteristics of the reference plant with the exception offive or fewer traits, 4 or fewer traits, 3 or fewer traits, 2 or fewertraits, or one trait. In embodiments, the plant comprising “essentiallyall of the physiological and morphological characteristics” of a variety(e.g., for tomato hybrid 603345), means a plant having all of thecharacteristics of the variety (e.g., mini-clusters of small maturefruit with a pale red color, a round shape, and/or a high fructosecontent or any other trait described herein, e.g., in Table 1).

“First water date”. The date the seed first receives adequate moistureto germinate. This can and often does equal the planting date.

“Gene”. As used herein, “gene” refers to a segment of nucleic acidcomprising an open reading frame. A gene can be introduced into a genomeof a species, whether from a different species or from the same species,using transformation or various breeding methods.

“Gene editing”: “Gene editing” (sometimes also called “genome editing”)refers to the introduction of targeted modifications into genomic DNAusing techniques employing, for example, meganucleases, zinc-fingernucleases, TALENs, and CRISPR/Cas9 systems. See, e.g., Gaj et al., ZFN,TALEN, and CRISPR/Cas-based methods for genome engineering. Trends inBiotechnology, 31:397-405 (2013).

“Inbred line”: As used herein, the phrase “inbred line” refers to agenetically homozygous or nearly homozygous population. An inbred line,for example, can be derived through several cycles of sib crossingand/or selfing and/or via double haploid production. In someembodiments, inbred lines breed true for one or more traits of interest.An “inbred plant” or “inbred progeny” is an individual sampled from aninbred line.

“Indeterminate”: Indeterminate tomato plants tend to be vine-like, andwill grow and produce fruits throughout the season (e.g., until killedby frost)

“Plant.” As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which plants can beregenerated, plant calli, plant clumps, and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos,cotyledons, hypocotyl, roots, root tips, anthers, pistils, flowers,ovules, seeds, fruit, stems, and the like.

“Plant material”. The terms “plant material” and “material obtainablefrom a plant” are used interchangeably herein and refer to any plantmaterial obtainable from a plant including without limitation, leaves,stems, roots, flowers or flower parts, fruits, pollen, ovules, zygotes,seeds, cuttings, cell or tissue cultures, or any other part or productof the plant.

“Plant part”. As used herein, a “plant part” includes any part, organ,tissue or cell of a plant including without limitation an embryo,meristem, leaf, pollen, cotyledon, hypocotyl, root, root tip, anther,flower, flower bud, pistil, ovule, seed, shoot, stem, stalk, petiole,pith, capsule, a scion, a rootstock and/or a fruit including callus andprotoplasts derived from any of the foregoing. In representativeembodiments, the plant part is a non-propagating plant part, forexample, is not a seed.

“Quantitative Trait Locus”. Quantitative Trait Locus (QTL) refers to agenetic locus that control to some degree, numerically representabletraits that are usually continuously distributed.

“Regeneration”. Regeneration refers to the development of a plant fromtissue culture.

“Resistance”. As used herein the terms “resistance” and “tolerance” (andgrammatical variations thereof) are used interchangeably to describeplants that show reduced or essentially no symptoms to a specific biotic(e.g., a pest, pathogen or disease) or abiotic (e.g., exogenous orenvironmental, including herbicides) factor or stressor. In someembodiments, “resistant” or “tolerant” plants show some symptoms but arestill able to produce marketable product with an acceptable yield, e.g.,the yield may still be reduced and/or the plants may be stunted ascompared with the yield or growth in the absence of the biotic and/orabiotic factor or stressor. Those skilled in the art will appreciatethat the degree of resistance or tolerance may be assessed with respectto a plurality or even an entire field of plants. A tomato plant may beconsidered “resistant” or “tolerant” if resistance/tolerance is observedover a plurality of plants (e.g., an average), even if particularindividual plants may be susceptible to the biotic or abiotic factor orstressor.

“RHS”. RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd., RHS Garden;Wisley, Woking; Surrey GU236QB, UK.

“Single locus conversion”. A single locus converted or conversion plantrefers to a plant that is developed by plant breeding techniques (e.g.,backcrossing), genome editing techniques, genetic transformationtechniques and/or mutation techniques wherein essentially all of thedesired morphological and physiological characteristics of a line arerecovered in addition to the single locus introduced into the line viathe plant breeding, genome editing, genetic transformation, or mutationtechniques.

“Substantially equivalent characteristic”. A characteristic that, whencompared, does not show a statistically significant difference (e.g.,p=0.05) from the mean.

“Transgene”. A nucleic acid of interest that can be introduced into thegenome of a plant by genetic engineering techniques (e.g.,transformation) or breeding. The transgene can be from the same or adifferent species. If from the same species, the transgene can be anadditional copy of a native coding sequence or can present the nativesequence in a form or context (e.g., different genomic location and/orin operable association with exogenous regulatory elements such as apromoter) than is found in the native state. The transgene can comprisean open reading frame encoding a polypeptide or can encode a functionalnon-translated RNA (e.g., an interfering RNA [RNAi]).

Botanical Description of Tomato Hybrid 603345.

When grown under standard horticultural conditions (e.g., in agreenhouse), the mature fruit of hybrid tomato 603345 is characterizedby a number of traits including, without limitation: a pale red maturefruit color, round shape, an average weight of about 24 grams, and ahigh fructose content.

More detailed botanical descriptions of hybrid 603345 are shown in Table1 below.

TABLE 1 Variety Description Information (based on evaluation in apassive greenhouse). Comparison of hybrid 603345 with a similar variety.Characteristic State of in which the State of expression of Denominationof similar variety expression of candidate similar variety is differentsimilar variety variety (603345) Nebula Fruit size Small to very Verysmall small (25 grams) (20 grams) Nebula Hot setting Susceptible Mediumsusceptibility

Additional information for tomato hybrid 603345.

Tissue Culture. Characteristic 603345 Plant: growth type IndeterminateTime of maturity (±comparable with the variety Late Katalina) Level ofsetting Medium-strong Seedling: anthocyanin coloration of hypocotylPresent Plant: height Medium-high Vigor of the plant Medium-strong Leaf:type of blade Bipinnate Leaf: length Medium Leaf: attitude (in middlethird of plant) Horizontal Leaf: intensity of green color Light-mediumDensity of foliage Medium Peduncle: abscission layer Present (jointed)Fruit: size Small Fruit: average weight in grams 24 Fruit: ratiolength/diameter Medium Fruit: shape in longitudinal section CircularFruit: cross section Round Fruit: size peduncle scar Very small-smallFruit: shape at blossom end Flat Fruit: shape of pistil scar PointedFruit: number of locules Two to three Fruit: green shoulder (beforematurity) Present Fruit: green stripes (before maturity) Absent Fruit:intensity of green color before maturity Light-medium Fruit: color atmaturity Red Fruit: firmness Firm-very firm Fruit: ribbing at peduncleend Absent Fruit: shelf life Medium-long Fruit: shelf life in days 12Homogeneity of size of the fruits Homogenous Long shelf life gene(s)present? Present Type of Long Shelf Life gene NOR Resistances: Fulviafulva group A Highly resistant Fulvia fulva group B Highly resistantFulvia fulva group C Highly resistant Fulvia fulva group D Highlyresistant Fulvia fulva group E Highly resistant Fusarium oxysporum f.sp. lycopersici race 0 Highly resistant Fusarium oxysporum f. sp.lycopersici race 1 Highly resistant Fusarium oxysporum f. sp.radicis-lycopersici Highly resistant Verticillium dahliae Highlyresistant Meloidogyne arenaria Intermediate resistant Meloidogyneincognita Intermediate resistant Meloidogyne javanica Intermediateresistant Tobacco Mosaic Virus race 0 Unknown Tomato Mosaic Virus strain0 Highly resistant Tomato Mosaic Virus strain 1 Highly resistant TomatoMosaic Virus strain 1.2 Highly resistant Tomato Mosaic Virus strain 2Highly resistant Tomato Spotted Wilt Virus Susceptible Tomato YellowLeaf Curl Virus Intermediate resistant Sensitivity to silvering Unknown

In embodiments, tomato plants can be propagated by tissue culture andregeneration. Tissue culture of various plant tissues and regenerationof plants therefrom is well known. For example, reference may be had toTeng, et al., HortScience, 27:9, 1030-1032 (1992); Teng, et al.,HortScience, 28:6, 669-1671 (1993); Zhang, et al., Journal of Geneticsand Breeding, 46:3, 287-290 (1992); Webb, et al., Plant Cell Tissue andOrgan Culture, 38:1, 77-79 (1994); Curtis, et al., Journal ofExperimental Botany, 45:279, 1441-1449 (1994); Nagata, et al., Journalfor the American Society for Horticultural Science, 125:6, 669-672(2000); and Ibrahim, et al., Plant Cell Tissue and Organ Culture, 28(2),139-145 (1992). It is clear from the literature that the state of theart is such that these methods of obtaining plants are routinely usedand have a high rate of success. Thus, another aspect of this inventionis to provide cells which upon growth and differentiation produce tomatoplants having desired characteristics of tomato hybrid 603345.Optionally, tomato plants can be regenerated from the tissue culture ofthe invention comprising all or essentially all of the physiological andmorphological characteristics of tomato hybrid 603345.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers, and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234, and5,977,445 describe certain techniques.

Additional Breeding Methods.

This invention is also directed to methods for producing a tomato plantby crossing a first parent tomato plant with a second parent tomatoplant wherein the first and/or second parent tomato plant is a plant oftomato hybrid 603345. Thus, any of the following exemplary methods usingtomato hybrid 603345 are part of this invention: selfing, backcrosses,hybrid production, crosses to populations, double haploid production,and the like. All plants produced using tomato hybrid 603345 as at leastone parent are within the scope of this invention, including thosedeveloped from tomato plants derived from tomato hybrid 603345.Advantageously, tomato hybrid 603345 can be used in crosses with other,different, tomato plants (e.g., inbred lines) to produce firstgeneration (F₁) tomato hybrid seeds and plants with desirablecharacteristics. The tomato plants of the invention can also be used fortransformation where exogenous transgenes are introduced and expressedby the plants of the invention or for introduction of genetic changes bygene editing or mutagenesis. Genetic variants created either throughtraditional breeding methods, gene editing, mutagenesis ortransformation of the cultivars of the invention by any of a number ofprotocols known to those of skill in the art are intended to be withinthe scope of this invention.

The following describes exemplary breeding methods that may be used withtomato hybrid 603345 in the development of further tomato plants. Onesuch embodiment is a method for developing tomato hybrid 603345 progenytomato plants (e.g., a diploid progeny tomato plant) in a tomato plantbreeding program comprising: obtaining a plant, or a part thereof, oftomato hybrid 603345, utilizing said plant or plant part as a source ofbreeding material, and selecting a tomato hybrid 603345 progeny plantwith some, all or essentially all morphological and/or physiologicalcharacteristics of tomato hybrid 603345 (see, e.g., Table 1),respectively, optionally with the use of molecular markers. Inrepresentative embodiments, a progeny plant of tomato hybrid 603345 hasat least 1, 2, 3, 4 or more of the morphological and physiologicalcharacteristics of tomato hybrid 603345 (for example, producesmini-clusters of small mature fruit with a pale red color, a roundshape, and/or a high fructose content). In embodiments, a progeny plantof tomato hybrid 603345 comprises essentially all or even all of themorphological and physiological characteristics of tomato hybrid 603345,respectively so that said progeny tomato plant is not significantlydifferent for said traits than tomato hybrid 603345, respectively, asdetermined at the 5% significance level when grown in the sameenvironmental conditions; optionally, with the presence of one or moredesired added traits. Breeding steps that may be used in the breedingprogram include pedigree breeding, backcrossing, mutation breeding,recurrent selection and/or double haploid production. In conjunctionwith these steps, techniques such as RFLP-enhanced selection, geneticmarker enhanced selection (for example, SSR markers) and/or and themaking of double haploids may be utilized.

Another representative method involves producing a population of tomatohybrid 603345 progeny plants (e.g., diploid progeny plants), comprisingcrossing tomato hybrid 603345 with another tomato plant, therebyproducing a population of tomato plants that, on average, derives atleast 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles (i.e., TAC) fromtomato hybrid 603345, e.g., at least about 6.25%, 12.5%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or99% of the genetic complement of tomato hybrid 603345. One embodiment ofthis invention is the tomato plant produced by this method and that hasobtained at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles (i.e.,TAC) from tomato hybrid 603345, and optionally may be the result of abreeding process comprising one or two breeding crosses and one or moreof selfing, sibbing, backcrossing and/or double haploid techniques inany combination and any order. In embodiments, the breeding process doesnot include a breeding cross, and comprises selfing, sibbing,backcrossing and or double haploid technology.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plants to determine if there is or is notsignificant difference between the two traits expressed by those plants.For example, see Fehr and Walt, Principles of Cultivar Development, pp.261-286 (1987). Thus, in embodiments, the invention includes tomatohybrid 603345 progeny tomato plants characterized by e.g., 1, 2, 3, 4 ormore characteristics of tomato hybrid 603345 as described herein, e.g.,produces mini-clusters of small mature fruit with a pale red color, around shape, and/or a high fructose content. In embodiments, theselected progeny comprises all or essentially all the morphological andphysiological characteristics of tomato hybrid 603345 (e.g., a describedin Table 1). In embodiments, the invention encompasses progeny plantshaving a combination of at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more ofthe characteristics as described herein for tomato hybrid 603345 (see,e.g., Table 1), so that said progeny tomato plant is not significantlydifferent for said traits than tomato hybrid 603345, respectively, asdetermined at the 5% significance level when grown in the sameenvironmental conditions. Using techniques described herein and thoseknown in the art, molecular markers may be used to identify said progenyplant as progeny of tomato hybrid 603345. Mean trait values may be usedto determine whether trait differences are significant, and optionallythe traits are measured on plants grown under the same environmentalconditions.

Progeny of tomato hybrid 603345 may also be characterized through theirfilial relationship with tomato hybrid 603345, as for example, beingwithin a certain number of breeding crosses of tomato hybrid 603345. Abreeding cross is a cross made to introduce new genetics into theprogeny, and is distinguished from a cross, such as a self or a sibcross or a backcross to a recurrent parent, made to select amongexisting genetic alleles. The lower the number of breeding crosses inthe pedigree, the closer the relationship between tomato hybrid 603345and its progeny. For example, progeny produced by the methods describedherein may be within 1, 2, 3, 4, 5 or more breeding crosses of tomatohybrid 603345.

In representative embodiments, a tomato plant derived from tomato hybrid603345 comprises cells comprising at least one set of chromosomesderived from tomato hybrid 603345. In embodiments, the tomato plant orpopulation of tomato plants derived from tomato hybrid 603345 comprises,on average, at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles(i.e., TAC) from tomato hybrid 603345, e.g., at least about 6.25%,12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%,95%, 96%, 97%, 98% or 99% of the genetic complement of tomato hybrid603345, and optionally may be the result of one or more of selfing,sibbing, backcrossing and/or double haploid techniques in anycombination. In embodiments, the tomato plant derived from tomato hybrid603345 is one, two, three, four, five or more breeding crosses removedfrom tomato hybrid 603345.

In representative embodiments, a plant derived from tomato hybrid 603345is a double haploid plant, a hybrid plant, an inbred plant, and/or adiploid plant.

In embodiments, a derived plant from tomato hybrid 603345 comprises adesired added trait. In representative embodiments, a tomato plantderived from tomato hybrid 603345 comprises all of the morphological andphysiological characteristics of tomato hybrid 603345 (e.g., asdescribed in Table 1). In embodiments, the tomato plant derived fromtomato hybrid 603345 comprises essentially all of the morphological andphysiological characteristics of tomato hybrid 603345 (e.g., asdescribed in Table 1), with the addition of a desired added trait.

Those skilled in the art will appreciate that traits can be introducedinto tomato hybrid 603345 by any method known in the art, e.g., planttransformation methods, conventional breeding, gene editing and/ornatural or induced mutations.

Further Embodiments of the Invention

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign nucleic acids including additional or modified versions ofnative (endogenous) nucleic acids (optionally driven by a non-nativepromoter) in order to alter the traits of a plant in a specific manner.Any nucleic acid sequences, whether from a different species, the samespecies or an artificial sequence, which are introduced into the genomeusing transformation or various breeding methods, are referred to hereincollectively as “transgenes.” Over the last fifteen to twenty years,several methods for producing transgenic plants have been developed, andin particular embodiments the present invention also relates totransformed versions of tomato plants disclosed herein.

Genetic engineering techniques can be used (alone or in combination withbreeding methods) to introduce one or more desired added traits intoplant, for example, tomato hybrid 603345 or progeny or tomato plantsderived thereof. Once a transgene has been introduction into a plant bygenetic transformation, it can be transferred to other plants viaconventional breeding.

Plant transformation generally involves the construction of anexpression vector that will function in plant cells. Optionally, such avector comprises one or more nucleic acids comprising a coding sequencefor a polypeptide or an untranslated functional RNA under control of, oroperatively linked to, a regulatory element (for example, a promoter).In representative embodiments, the vector(s) may be in the form of aplasmid, and can be used alone or in combination with other plasmids, toprovide transformed tomato plants using transformation methods asdescribed herein to incorporate transgenes into the genetic material ofthe tomato plant.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct nucleic acid transfermethod, such as microprojectile-mediated delivery (e.g., with abiolistic device), DNA injection, Agrobacterium-mediated transformation,electroporation, and the like. Transformed plants obtained from theplants (and parts and tissue culture thereof) of the invention areintended to be within the scope of this invention.

Expression Vectors for Plant Transformation—Selectable Markers.

Expression vectors typically include at least one nucleic acidcomprising or encoding a selectable marker, operably linked to aregulatory element (for example, a promoter) that allows transformedcells containing the marker to be either recovered by negativeselection, e.g., inhibiting growth of cells that do not contain theselectable marker, or by positive selection, e.g., screening for theproduct encoded by the selectable marker. Many commonly used selectablemarkers for plant transformation are well known in the transformationart, and include, for example, nucleic acids that code for enzymes thatmetabolically detoxify a selective chemical agent which may be anantibiotic or an herbicide, or nucleic acids that encode an alteredtarget which is insensitive to the inhibitor. Positive selection methodsare also known in the art.

Commonly used selectable markers in plants include, but are not limitedto: neomycin phosphotransferase II (nptII) conferring resistance tokanamycin, hygromycin phosphotransferase conferring resistance to theantibiotic hygromycin, bacterial selectable markers that conferresistance to antibiotics (e.g., gentamycin acetyl transferase,streptomycin phosphotransferase, and aminoglycoside-3′-adenyltransferase, selectable markers conferring resistance to herbicides(e.g., glyphosate, glufosinate, or bromoxynil). Selection of transformedplant cells can also be based on screening presumptively transformedplant cells rather than direct genetic selection of transformed cellsfor resistance to a toxic substance such as an antibiotic; such markersinclude without limitation alpha-glucuronidase (GUS),alpha-galactosidase, luciferase, and Green Fluorescent Protein (GFP) andmutant GFPs.

Expression Vectors for Plant Transformation—Promoters.

Transgenes included in expression vectors are generally driven by anucleotide sequence comprising a regulatory element (for example, apromoter). Numerous types of promoters are well known in thetransformation arts, as are other regulatory elements that can be usedalone or in combination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells.

Examples of promoters under developmental control include promoters thatpreferentially initiate transcription in certain tissues, such asleaves, roots, seeds, fibers, xylem vessels, tracheids, or sclerenchyma.Such promoters are referred to as “tissue-preferred.” Promoters thatinitiate transcription only in certain tissue are referred to as“tissue-specific.” A “cell type” specific promoter preferentially drivesexpression in certain cell types in one or more organs, for example,vascular cells in roots or leaves. An “inducible” promoter is a promoterthat is under environmental control. Examples of environmentalconditions that may affect transcription by inducible promoters includeanaerobic conditions or the presence of light. Tissue-specific,tissue-preferred, cell type specific, and inducible promoters constitutethe class of “non-constitutive” promoters. A “constitutive” promoter isa promoter that is active under most environmental conditions.

Many suitable promoters are known in the art and can be selected andused to achieve the desired outcome.

Signal Sequences for Targeting Proteins to Subcellular Compartments.

Transport of polypeptides produced by transgenes to a subcellularcompartment such as the chloroplast, vacuole, peroxisome, glyoxysome,cell wall, or mitochondrion, or for secretion into the apoplast, isgenerally accomplished by means of operably linking a nucleotidesequence encoding a signal sequence to the 5′ and/or 3′ region of anucleic acid encoding the polypeptide of interest. Signal sequences atthe 5′ and/or 3′ end of the coding sequence target the polypeptide toparticular subcellular compartments.

The presence of a signal sequence can direct a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample, Becker, et al., Plant Mol. Biol., 20:49 (1992); Close, P. S.,Master's Thesis, Iowa State University (1993); Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol., 9:3-17 (1987); Lerner, et al., PlantPhysiol., 91:124-129 (1989); Fontes, et al., Plant Cell, 3:483-496(1991); Matsuoka, et al., PNAS, 88:834 (1991); Gould, et al., J. Cell.Biol., 108:1657 (1989); Creissen, et al., Plant J, 2:129 (1991);Kalderon, et al., A short amino acid sequence able to specify nuclearlocation, Cell, 39:499-509 (1984); and Steifel, et al., Expression of amaize cell wall hydroxyproline-rich glycoprotein gene in early leaf androot vascular differentiation, Plant Cell, 2:785-793 (1990).

Foreign Polypeptide Transgenes and Agronomic Transgenes.

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign polypeptide then canbe extracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem., 114:92-6(1981). According to a representative embodiment, the transgenic plantprovided for commercial production of foreign protein is a tomato plantof the invention. In another embodiment, the biomass of interest is seedand/or fruit.

Likewise, by means of the present invention, agronomic transgenes andother desired added traits can be expressed in transformed plants (andtheir progeny, e.g., produced by breeding methods). More particularly,plants can be genetically engineered to express various phenotypes ofagronomic interest or other desired added traits. Exemplary nucleicacids of interest in this regard conferring a desired added trait(s)include, but are not limited to, those transgenes that confer resistanceto confer resistance to plant pests (e.g., nematode or insect) ordisease (e.g., fungal, bacterial or viral), transgenes that conferherbicide tolerance, transgenes that confer male sterility, andtransgenes that confer or contribute to a value-added trait such asincreased nutrient content (e.g., iron, nitrate), increased sweetness(e.g., by introducing a transgene coding for monellin), modified fattyacid metabolism (for example, by introducing into a plant an antisensesequence directed against stearyl-ACP desaturase to increase stearicacid content of the plant), modified carbohydrate composition (e.g., byintroducing into plants a transgene coding for an enzyme that alters thebranching pattern of starch), modified fruit color (e.g., external fruitcolor and/or fruit flesh), or modified flavor profile of the fruit.

In embodiments, the transgene encodes a non-translated RNA (e.g., RNAi)that is expressed to produce targeted inhibition of gene expression,thereby conferring the desired trait on the plant.

In embodiments, the transgene encodes the machinery used for geneediting techniques.

Any transgene, including those exemplified above, can be introduced intothe tomato plants of the invention through a variety of means including,but not limited to, transformation (e.g., genetic engineeringtechniques), conventional breeding, and introgression methods tointroduce the transgene into other genetic backgrounds.

Methods for Plant Transformation.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glickand Thompson Eds., CRC Press, Inc., Boca Raton, pp. 67-88 (1993). Inaddition, expression vectors and in vitro culture methods for plant cellor tissue transformation and regeneration of plants are available. See,for example, Gruber, et al., “Vectors for Plant Transformation” inMethods in Plant Molecular Biology and Biotechnology, Glick and ThompsonEds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993). Commonly usedplant transformation methods include agrobacterium-mediatedtransformation and direct transgene transfer methods (e.g.,microprojectile-mediated transformation, sonication, liposome orspheroplast fusion, and electroporation of protoplasts or whole cells).

Following transformation of plant target tissues, expression ofselectable marker transgenes (e.g., as described above) allows forpreferential selection of transformed cells, tissues and/or plants,using regeneration and selection methods now well known in the art.

The foregoing methods for transformation are typically used to produce atransgenic tomato line. The transgenic tomato line can then be crossedwith another (non-transgenic or transgenic) line in order to produce anew transgenic tomato line. Alternatively, a transgene that has beenengineered into a particular plant using transformation techniques canbe introduced into another plant or line using traditional breeding(e.g., backcrossing) techniques that are well known in the plantbreeding arts. For example, a backcrossing approach can be used to movean engineered transgene from a public, non-elite inbred line into anelite inbred line, or from an inbred line containing a foreign transgenein its genome into an inbred line or lines which do not contain thattransgene. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context.

Locus Conversions.

When the term “plant” is used in the context of the present invention,this term also includes any locus conversions of that plant or variety.The term “locus converted plant” as used herein refers to those plantsthat are developed, for example, by backcrossing, genome editing,genetic transformation and/or mutation, wherein essentially all of thedesired morphological and physiological characteristics of a variety(e.g., for tomato hybrid 603345, produces mini-clusters of small maturefruit with a pale red color, a round shape, and/or high fructose contentor any other trait described herein, e.g., in Table 1) are recovered inaddition to the one or more genes introduced into the variety.

To illustrate, backcrossing methods can be used with the presentinvention to improve or introduce a characteristic into the variety. Theterm “backcrossing” as used herein refers to the repeated crossing of ahybrid progeny back to the recurrent parent, e.g., backcrossing 1, 2, 3,4, 5, 6, 7, 8, 9, or more times to the recurrent parent. The parentalplant that contributes the gene for the desired characteristic is termedthe “nonrecurrent” or “donor parent.” This terminology refers to thefact that the nonrecurrent parent is generally used one time in thebreeding e.g., backcross) protocol and therefore does not recur. Thegene that is transferred can be a native gene, a mutated native gene ora transgene introduced by genetic engineering techniques into the plant(or ancestor thereof). The parental plant into which the gene(s) fromthe nonrecurrent parent are transferred is known as the “recurrent”parent as it is used for multiple rounds in the backcrossing protocol.Poehlman & Sleper (1994) and Fehr (1993). In a typical backcrossprotocol, the original variety of interest (recurrent parent) is crossedto a second variety (nonrecurrent parent) that carries the gene(s) ofinterest to be transferred. The resulting progeny from this cross arethen crossed again to the recurrent parent and the process is repeateduntil a plant is obtained wherein essentially all of the desiredmorphological and physiological characteristics of the recurrent parentare recovered in the converted plant in addition to the transferredgene(s) and associated trait(s) from the nonrecurrent parent.

Genetic Analysis of Tomato Hybrid 603345.

The invention further provides a method of determining a geneticcharacteristic of tomato hybrid 603345 or a progeny thereof, e.g., amethod of determining a genotype of tomato hybrid 603345 or a progenythereof. In embodiments, the method comprises detecting in the genome ofa tomato hybrid 603345 plant, or a progeny plant thereof, at least afirst polymorphism (e.g., by detecting a nucleic acid marker by a methodcomprising nucleic acid amplification and/or nucleic acid sequencing).To illustrate, in embodiments, the method comprises obtaining a sampleof nucleic acids from the plant and detecting at least a firstpolymorphism in the nucleic acid sample. Optionally, the method maycomprise detecting a plurality of polymorphisms (e.g., two or more,three or more, four or more, five or more, six or more, eight or more orten or more polymorphisms, etc.) in the genome of the plant. Inrepresentative embodiments, the method further comprises storing theresults of the step of detecting the polymorphism(s) on a computerreadable medium. The invention further provides a computer readablemedium produced by such a method.

Although the present invention has been described with reference tospecific details of certain embodiments thereof, it is not intended thatsuch details should be regarded as limitations upon the scope of theclaimed invention except as and to the extent that they are included inthe accompanying claims.

DEPOSIT INFORMATION

Applicants have made a deposit of at least 2500 seeds of tomato hybridOnuba with American Type Culture Collection (ATCC), 10801 UniversityBoulevard, Manassas, Va., 20110-2209 U.S.A. ATCC under Accession No.PTA-127138 on Dec. 13, 2021 under the provisions of the Budapest Treaty.The deposit of tomato hybrid Onuba will be maintained in the ATCCdepository, which is a public depository, for a period of 30 years, or 5years after the most recent request, or for the effective life of thepatent, whichever is longer, and will be replaced if any of thedeposited seed becomes nonviable during that period. Additionally,Applicants have satisfied all the requirements of 37 C.F.R. § §1.801-1.809, including providing an indication of the viability of thesamples. Access to these deposits will be made available during thependency of this application to the Commissioner upon request. Allrestrictions on the availability of the deposited material from ATCC tothe public will be irrevocably and without restriction removed upongranting of the patent. Applicants impose no restrictions on theavailability of the deposited material from ATCC; however, Applicantshave no authority to waive any restrictions imposed by law on thetransfer of biological material or its transportation in commerce.Applicants do not waive any infringement of its rights granted underthis patent or under the Plant Variety Protection Act (7 USC § 2321 etseq.).

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be apparent that certain changes and modifications suchas single locus modifications and mutations, somaclonal variants,variant individuals selected from large populations of the plants of theinstant cultivars and the like may be practiced within the scope of theinvention.

What is claimed is:
 1. A seed that produces tomato hybrid Onuba, arepresentative sample of seed having been deposited under ATCC AccessionNo. PTA-127138.
 2. A plant of tomato hybrid Onuba produced from the seedof claim
 1. 3. A tomato plant, or a part thereof, having all thephysiological and morphological characteristics of the tomato plant ofclaim
 2. 4. A seed that produces the plant of claim
 3. 5. A plant partof the plant of claim
 2. 6. The plant part of claim 5, wherein the plantpart is a leaf, a fruit, a shoot, pollen, an ovule, an anther, a root, arootstock, a scion, or a cell.
 7. A tissue culture of regenerable cellsof the tomato plant of claim
 2. 8. A tomato plant regenerated from thetissue culture of claim 7, wherein said tomato plant comprises all ofthe physiological and morphological characteristics of tomato hybridOnuba.
 9. A method of producing tomato seed, the method comprisingcrossing the plant of claim 2 with itself or a second tomato plant andharvesting the resulting seed.
 10. A grafted tomato plant comprising arootstock and a scion, wherein the plant of claim 2 is used as the scionand the rootstock is from a different tomato plant.
 11. A method ofproducing a grafted tomato plant, the method comprising: (a) providing ascion from the plant of claim 2; and (b) grafting the scion to arootstock from a different tomato plant.
 12. A method of developing atomato line in a tomato plant breeding program using plant breedingtechniques, which include employing a tomato plant, or a part thereof,as a source of plant breeding material, the method comprising: (a)obtaining the tomato plant, or part thereof, of claim 2 as a source ofbreeding material; and (b) applying plant breeding techniques to saidtomato plant or part thereof.
 13. A method for producing a seed of atomato plant derived from the tomato hybrid Onuba, the methodcomprising: (a) crossing the plant of claim 2 with a different tomatoplant; (b) allowing seed to form; (c) growing a plant from the seed ofstep (b) to produce a plant derived from tomato hybrid Onuba; (d)selfing the plant of step (c) or crossing it to a second tomato plant toform additional tomato seed derived from tomato hybrid Onuba; and (e)optionally repeating steps (c) and (d) one or more times to generatefurther derived tomato seed from tomato hybrid Onuba, wherein in step(c) a plant is grown from the additional tomato seed of step (d) inplace of growing a plant from the seed of step (b).
 14. A method ofvegetatively propagating tomato hybrid Onuba, the method comprising: (a)collecting tissue capable of being propagated from the plant of claim 2;(b) cultivating the tissue to obtain proliferated shoots; (c) rootingthe proliferated shoots to obtain rooted plantlets; and (d) optionally,growing plants from the rooted plantlets.
 15. A tomato plantlet or plantobtained by the method of claim 14, wherein the tomato plantlet or plantcomprises all of the physiological and morphological characteristics oftomato hybrid Onuba.
 16. A method of producing a plant of tomato hybridOnuba comprising a desired added trait, the method comprisingintroducing a transgene conferring the desired trait into the plant ofclaim
 2. 17. A tomato plant produced by the method of claim 16, whereinthe tomato plant comprises the transgene conferring the desired traitand otherwise comprises all of the morphological and physiologicalcharacteristics of tomato hybrid Onuba.
 18. A seed that produces theplant of claim
 17. 19. A method for producing a tomato seed, the methodcomprising selfing the plant of claim 2 for one or more generations andallowing seed to form.
 20. A method of producing a tomato fruit, themethod comprising: (a) growing the tomato plant according to claim 2 toproduce a tomato fruit; and (b) harvesting the tomato fruit.
 21. Amethod of producing a processed tomato product, the method comprising:(a) obtaining a fruit of the plant of claim 2; and (b) processing saidfruit to produce a processed product.
 22. A method of determining agenotype of tomato hybrid Onuba, the method comprising: (a) obtaining asample of nucleic acids from the plant of claim 2; and (b) detecting apolymorphism in the nucleic acid sample using molecular biologytechniques, thereby determining the genotype of tomato hybrid Onuba.